Peroxin 14 Double Nickase Plasmid (h): sc-405075-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Peroxin 14 Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- Peroxin 14 Double Nickase Plasmid (h) and Peroxin 14 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PEX14. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Peroxin 14 Antibody (1G12): sc-293383
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Peroxin 14 Double Nickase Plasmid (h) | sc-405075-NIC | 20 µg | $410.00 | |||
Peroxin 14 Double Nickase Plasmid (h2) | sc-405075-NIC-2 | 20 µg | $410.00 |
PEX14 encodes peroxin 14, an integral component of the peroxisomal membrane that functions as a docking factor for the PEX5 and PEX7 receptor complexes during import of matrix enzymes bearing PTS1 and PTS2 targeting signals. Through this role, PEX14 is central to peroxisome biogenesis, maintenance of organelle proteostasis, and metabolic pathways including very-long-chain fatty acid β-oxidation and cellular redox homeostasis. Perturbation of PEX14-dependent import disrupts peroxisomal metabolism and can secondarily affect mitochondrial function, lipid signaling, and reactive oxygen species handling. Dysregulation of peroxisome assembly and import machinery is implicated in peroxisome biogenesis disorders and broader mechanisms relevant to neurodevelopment and metabolic dysfunction.
Peroxin 14 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PEX14 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PEX14. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt PEX14 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of PEX14-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.